抗致病性大肠杆菌O_(78)特异单克隆抗体的研制及其初步应用

Monclonal Antibodies with Specificity to Avain pathoigenic E. coli. O_(78) and Their potential uses as Disagnostic Reagents

应用淋巴细胞杂交瘤技术将提纯O_78:K_80：H_9标准菌株免疫BALB／C小鼠的脾细胞与小鼠骨髓瘤细胞融合，获得5株能稳定分泌抗体的杂交瘤细胞命名为1G_7、2C_6、3C_1、3E_1、3D_12，这5株杂交瘤细胞经三次克隆化都能稳定分泌抗体。玻板凝集试验、酶联免疫吸附试验和间接免疫荧光试验结果表明：这5株单抗对所试37个菌株中O_78菌株都能发生反应。但3E_1、3D_12还可与另外6个菌株出现交叉反应。3C_1可与另外所试14个菌株发生反应，1G_7和2C_6则只与O_78菌株发生反应。用上述1G_7，单抗对临床分离123株细菌进行鉴定，与常规单因子血清鉴定结果完全符合。试验结果表明对，O_78特异单抗可以取代常规O_78单因子血清。

A panel of five hybridoma cell lines, 1G_7, 2C_6, 3C_1, 3E_1, 3D_12 scereting monoclonal antibodies(McAb) were obtained by fusing mouse myeloma cell line SP2/0 - Ag_14 and spleen cells from BALB/C mice immunized with purified O_78,K_80: H_9. The antibodies from culture medium had titers of 1: 1 in the direct agglutination test (DA ),10 -102 in the indirect immynofluorescence test (IIF ), 10~102 in the indirect immunofluorescence test (IIF), 10- 102 in the enzyme - linked immuno - sorbent assay (ELISA ) and the antibodies from the ascitic fluid had titers of 10- 103 in DA, 104 - 106 in the IIF and 104 - 106 in the ELISA. The specificity of these antibodes was evaluated by assaying their binding reactivities to a set of 27 strains of E- coli., and 10 strains of other enteric bacteria. As shown by DA, IIF and ELISA, 1G7. 2C6 McAbs reacted only with strains bearing O_78,3E1, 3D12 McAbs reacted with strairs bearing O_78and other fourteen strains of E. coli. The specificities of these McAbs were further verified hy their reactivity patterns with a total of 123 field isolales, in which 118 were E.coli., the other 5 were unidentified bacteria. The results coincided well with conventional single - factor O_78 antiserum. Of the 123 isolates, 26 strains O_78were identified hy both monoclonal and polyclonal reagents